Skip to content
← Back to browse
PROSTATE Publication-derived

Organoid Cultures Derived from Patients with Advanced Prostate Cancer

Source Gao et al., 2014 · Memorial Sloan Kettering Cancer Center · 10.1016/j.cell.2014.08.016

👤 Dong Gao, Ian Vela, Andrea Sboner, Phillip J. Iaquinta, Wouter R. Karthaus, Anuradha Gopalan, Catherine Dowling, Jackline N. Wanjala, Eva A. Undvall, Vivek K. Arora, John Wongvipat, Myriam Kossai, Sinan Ramazanoglu, Luendreo P. Barboza, Wei Di, Zhen Cao, Qi Fan Zhang, Inna Sirota, Leili Ran, Theresa Y. MacDonald, Himisha Beltran, Juan-Miguel Mosquera, Karim A. Touijer, Peter T. Scardino, Vincent P. Laudone, Kristen R. Curtis, Dana E. Rathkopf, Michael J. Morris, Daniel C. Danila, Susan F. Slovin, Stephen B. Solomon, James A. Eastham, Ping Chi, Brett Carver, Mark A. Rubin, Howard I. Scher, Hans Clevers, Charles L. Sawyers, Yu Chen ⏱ 56 days 📋 10 phases 🧫 Patient-Derived Prostate Cancer (Advanced/Castration-Resistant)

Abstract

This protocol describes the establishment and culture of patient-derived prostate cancer organoid lines from metastatic biopsies and circulating tumor cells. Seven organoid lines were successfully derived and extensively characterized for genomic alterations, gene expression, and drug sensitivity, representing diverse prostate cancer subtypes including AR-dependent adenocarcinoma, AR-negative adenocarcinoma, neuroendocrine carcinoma, and squamous differentiation.

Cell source
Patient-Derived Prostate Cancer (Advanced/Castration-Resistant)
Application
Disease modeling, Drug screening, Patient-derived tumor models

Protocol overview

37 steps across 10 phases

Tissue Acquisition and Processing Day 0
  1. 1 Tissue Biopsy Collection
  2. 2 Mince Tissue Sample
  3. 3 Collagenase Digestion
  4. 4 Cell Washing
CTC Isolation (when applicable) Day 0
  1. 1 CD45 Depletion
  2. 2 Ficoll-Paque Separation
  3. 3 CTC Washing
Organoid Seeding and Culture Initiation Day 0–1
  1. 1 Matrigel Seeding
  2. 2 Culture in Prostate Organoid Medium
  3. 3 Monitor Organoid Formation
Organoid Passaging and Propagation Day 7 onwards (weekly passage)
  1. 1 Select for Continuous Propagation
  2. 2 Dissociate Organoids for Passaging
  3. 3 Passage Organoids
Genomic Characterization – Sample Preparation Upon establishment of stable lines
  1. 1 DNA Extraction
  2. 2 RNA Extraction
Whole-Exome Sequencing and Analysis Days 1–14 (sequencing and data processing)
  1. 1 Library Preparation
  2. 2 Paired-End Sequencing
  3. 3 Sequence Alignment
  4. 4 Quality Score Recalibration and Indel Realignment
  5. 5 Variant Calling
  6. 6 Somatic Mutation Filtering
RNA-Sequencing and Expression Analysis Days 1–14 (sequencing and data processing)
  1. 1 RNA Library Preparation and Sequencing
  2. 2 Read Alignment and Quantification
  3. 3 Clustering and Heatmap Analysis
Array CGH and Copy Number Analysis Days 1–7 (sample processing and analysis)
  1. 1 Array CGH Preparation
  2. 2 Segmentation and Copy Number Calling
In Vitro Drug Sensitivity Testing Days 1–4 (per assay)
  1. 1 Cell Plating for Viability Assay
  2. 2 Drug Treatment
  3. 3 Viability Measurement
  4. 4 IC50 Determination
In Vivo Xenograft Studies Days 0–56+ (tumor establishment and treatment)
  1. 1 Organoid Cell Preparation
  2. 2 Matrigel Mixing
  3. 3 Xenograft Implantation
  4. 4 Tumor Growth Monitoring
  5. 5 Drug Treatment Initiation
  6. 6 Tumor Response Assessment
  7. 7 Tissue Harvest and Histology

Full SOP

🔬

Create a free account to access this protocol

Join OrganMatch to unlock step-by-step procedures, reagent concentrations, QC checklists, and downloadable batch record templates.

Create free account

Already registered? Log in

Attribution

This SOP was authored by Organthis based on the published method in Gao et al., 2014. The originating laboratory holds no rights in this SOP and has not endorsed it unless marked Verified.

This wording is awaiting legal review.

Something wrong with this entry? Report an issue with this protocol

Need a commercial licence?
Use this protocol in your therapeutic or diagnostic pipeline.
Request Commercial Licence